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Follicular development after ovum pick-up and fertilizability of retrieved oocytes in postpartum dairy cattle.

This study aimed to evaluate gonadotropin secretion and the developmental competence of follicular oocytes in dairy cattle during the early postpartum (PP) period. The number of follicles developed after transvaginal ultrasound-guided ovum pick-up (OPU) and fertilizability of retrieved oocytes were compared between cows in which the first dominant follicle (DF) ovulated (ovulated group, n=4) and did not ovulate (non-ovulated group, n=3), and between early PP (early PP group, n=2) and after the resumption of the estrous cycle (cyclic group, n=2). Follicular ablation was performed 2-4 days after the detection of DF in the second follicular wave PP. OPU was repeated 3-5 times at 3 or 4-day intervals from 3-4 days after the follicular ablation. At OPU, the follicles were enumerated and all those > or = 5 mm in diameter were aspirated. Recovered oocytes were subjected to in vitro maturation and fertilization. Both criteria were similar between ovulated and non-ovulated groups, and between early PP and cyclic groups. These results suggest that FSH/LH secretions required for follicle recruitment and subsequent follicular growth during the early PP period are similar to those after resumption of the estrous cycle. They also indicate that follicular oocytes during the early PP period have developmental competence.

Animals↗

Follicular development, steroidogenesis and ovulation within ovaries exposed in vitro to hormone levels which mimic those of the rat estrous cycle.

In order to more clearly define the hormonal factors responsible for follicular growth, ovulation and steroidogenesis, a perifusion culture system was developed. This culture system was capable of exposing the ovaries to hormonal levels which mimic the 4-day estrous cycle of the rat. Using this culture system, ovaries were exposed to the in vivo levels of luteinizing hormone (LH), follicle-stimulating hormone (FSH), estradiol-17 beta and progesterone to within the mean +/- one standard deviation of the values reported by Butcher et al. (1974a). During the 4-day culture period, the diameter of the follicles gradually increased in a manner similar to that of cycling rats. After 4 days in vitro, the ovaries ovulated 4.3 +/- 0.6 oocytes/ovary as assessed by the presence of ruptured-partially luteinized follicles. This in vitro ovulation rate was not significantly different from the in vivo ovulation rate of 5.0 +/- 0.8 oocytes/ovary (P greater than 0.05). The pattern of follicular growth within cultured ovaries was similar to that of the cycling rat, although the number of small follicles recruited to grow was reduced. The steroid secretory patterns also were slightly different in the cultured ovaries. These data indicate that physiological ovarian responses which are similar to those of cycling rats can be induced within cultured ovaries. With further utilization of this culture technique, the precise role of gonadotropins, ovarian steroids and other hormonal agents can be examined in order to define specific ovarian functions which are controlled by each hormonal agent.

Animals↗

The dominant follicle exerts an interovarian inhibition on FSH-induced follicular development.

An experiment was conducted to evaluate the role of the dominant follicle (DF) of the first wave in regulating follicular and ovulatory responses and embryonic yield to a superovulation regime with FSH-P. Twenty normally cycling Holstein-Freisian heifers (n = 20) were synchronized with GnRH and pgf(2alpha) and randomly assigned to a control or a treated group (n = 10 each). Treated heifers had the first wave dominant follicle removed via transvaginal, ultrasound-guided aspiration on Day 6 after a synchronized estrus. All heifers received a total of 32 mg FSH-P given in decreasing doses at 12 h intervals from Day 8 to Day 11 plus two injections of pgf(2alpha) (35 mg and 20 mg, respectively) on Day 10. Heifers were inseminated at 6 h and 16 h after onset of estrus. Follicular dynamics were examined daily by transrectal ultrasonography from Day 4 to estrus, once following ovulation, and at the time of embryo collection on Day 7. Blood samples were collected daily during the superovulatory treatment and at embryo collection. Follicles were classified as: small, </= 5 mm; medium, 6-9 mm; or large, >/= 10 mm. Aspiration of the dominant follicle was associated with an immediate decrease in large follicles, and a linear rate increase in small follicles from Day 4 to Day 8 just prior to the FSH-P injections, (treatment > control: +0.33 vs. -0.22, number of small follicles per day; P < 0.10). During FSH-P injections, the increase in number of medium follicles was greater (P < 0.01) for treatment on Day 9-11 (treatment > control: Day 9, 3.2 > 1.8; Day 10, 9.2 > 4.7; Day 11, 13.1 > 8.3; +/- 0.56). Number of large follicles was greater in treatment at Day 11 (5.12 > 1.4 +/-0.21; P < 0.01). Mean number of induced ovulatory follicles (difference between number of follicles at estrus and Day 2 after estrus) was greater in treatment (13.4 > 6.3 +/- 1.82; P < 0.01). Plasma estradiol at Day 11 during FSH-P treatment was greater in treatment (32.5 > 15.8 +/- 2.6; P < 0.01). Plasma progesterone at embryo flushing (Day 7 after ovulation) was greater in treatment (7.4 > 4.9; P < 0.02); technical difficulties at embryo recovery reduced sensitivity of embryonic measurements. No changes in the distribution of unfertilized oocytes and embryo developmental stages were detected between control and treatment groups. Presence of dominant follicle of the first wave inhibited intraovarian follicular responses to exogenous FSH.

Journal Article↗

Overview of ovarian follicular development: considerations for the toxicologist.

Folliculogenesis is the lengthy process that results in the production of a species-specific, highly consistent number of follicles, which ripen during each reproductive cycle at precisely the appropriate time for ovulation. Certain features of folliculogenesis may have special implications for toxicologists studying effects of environmental mutagens on oocytes. Such features include the constantly changing geometry of the ovarian follicle, the great excess of developing follicles (most of which will degenerate rather than ovulate), the exponential nature of follicular growth, the acceleration of cell proliferation as follicular size increases, and the location of the principal feedback regulatory step at the penultimate stage of the developmental process. Because the ovary can respond quickly and completely to loss of homeostasis over the short term, damage from toxic insult may not be readily apparent. However, long-range fertility may nevertheless be impaired. The finite size of the follicular pool and the absence of feedback regulatory steps during the early stages of follicular growth render the ovary incapable of restoring the status quo among small and medium-sized follicles. This will eventually result in loss of fine control over the number of follicles that ripen and the regularity of the reproductive cycles and could reduce the overall duration of the fertile life span.

Animals↗

[Angiogenesis in reproductive physiology. Follicular development, formation and maintenance of the corpus luteum].

The process of new capillary formations from previously existing mature vessels in healthy individuals has been mainly studied during cycles of the female reproductive tract. This new capillary formation, known as angiogenesis is related to endogenous regulators that both stimulates or inhibits it. Knowledge on the role of both stimulators and inhibitors under physiological and pathological conditions accentuates a main role of VEGF, FGF, angiogenin and angiopoietics among the formers; and main role of angiostatin and endostatin among the latters. In recent years, angiogenesis in the ovaries that leads to follicular and luteal growth and development has been extensively studied. Whether a number of endogenous stimulators and inhibitors have been identified, the molecular link between the endocrine and vascular system is not fully understood. Therefore, efforts to formulate questions that need answers must be made.

Corpus Luteum↗

Inhibin and follicular development in heterotopical ovary transplants without vascular pedicle in syngeneic Lewis rats.

OBJECTIVE: To evaluate the role of inhibin in elevated base levels of FSH and follicular hyperplasia in ovarian autotransplantation in rats. DESIGN: Experimental animal study. SETTING: Unit of Experimental Research at the Barcelona University School of Medicine. ANIMAL(S): Female syngeneic Lewis rats aged 16 weeks. INTERVENTION(S): The animals were randomized into two groups: group A, control group undergoing only laparotomy (n = 5) and group B, oophorectomized with SC autologous heterotopic transplant (n = 5). The animals were killed and their ovaries removed for histologic, morphometric, and immunohistochemical analysis at 28 days after surgery in both groups. MAIN OUTCOME MEASURE(S): Serum levels of E2 and FSH were determined on day 0 (the day of surgery or baseline) and days 4, 7, 14, 21, and 28. Morphometric analysis of ovarian structure for evaluation of antral follicles and their granulosa cell area and immunohistochemistry for inhibin staining were also done. RESULT(S): The endocrinological function recovered at 28 days, and the FSH levels for the transplant group were significantly higher than for the group with normoinsert ovary. Morphometric analysis showed that the mean granulosa cell area was greater in group B when compared with the control group. Immunohistochemistry revealed almost null inhibin staining of the stroma in transplanted ovarian tissue. CONCLUSION(S): Tissue damage brought on by ischemia in the transplant of nonvascularized ovaries may bring about an inhibin deficit in the ovarian stroma, which might explain the increased levels of FSH. These increased levels, in turn, would be responsible for the follicular hyperplasia seen in this tissue when it recovers its function.

Animals↗

Ortho Evra/Evra versus oral contraceptives: follicular development and ovulation in normal cycles and after an intentional dosing error.

OBJECTIVE: To compare the effects of the contraceptive patch to oral contraceptives (OCs) on follicular size and incidence of ovulation in normal cycles and after dosing errors. DESIGN: Randomized, open-label. SETTING: Twelve centers. PATIENT(S): One hundred twenty-four ovulatory women. INTERVENTION(S): Subjects received either the patch (groups 1 and 2) or one of three OCs. Correct dosing occurred in cycles 1, 2, 3, and 5. The following dosing errors were planned during cycle 4, a shortened 10-day cycle: [1] patch group 1 subjects wore one patch for 10 consecutive days; [2] for patch group 2 and OC subjects, 7 dosing days were followed by 3 drug-free days. MAIN OUTCOME MEASURE(S): Follicular size, as determined at each cycle by the maximum mean follicular diameter. RESULT(S): After a 3-day dosing error, follicular size was significantly smaller in the patch group (mean, 7.0 mm) vs. each OC group (range of means, 11.8-17.1 mm). Similar results were seen after proper dosing. The incidence of ovulation was significantly lower for the patch users than for women using OCs. CONCLUSION(S): Follicular size and incidence of ovulation were significantly reduced among contraceptive patch users compared with women using OCs in normal cycles and after planned dosing errors.

Administration, Cutaneous↗

Endocrine profiles and follicular development in early-weaned postpartum beef cows.

This study investigated whether treatment with progestagen, which improves fertility after early weaning in postpartum cows, altered concentrations of gonadotrophins or development and function of follicles. Patterns of luteinizing hormone (LH), oestradiol and follicle-stimulating hormone (FSH) and of follicular growth before first postpartum ovulation were compared in two experiments. At 17 to 25 days post partum, suckled anoestrous beef cows received an ear-implant containing 6 mg progestagen (norgestomet) for 9 days or served as untreated controls. Calves were weaned from all cows 7 days after initiation of treatment. Cows were observed for oestrous behaviour twice a day until 10 days after weaning. As expected, the proportion of anoestrous cows that formed a corpus luteum with a normal lifespan was greater (P < 0.01) in cows treated with norgestomet (Expt 1, 17 of 24; Expt 2, 18 of 22) than in control cows (Expt 1, 2 of 16; Expt 2, 3 of 18). In general, patterns of secretion of LH and oestradiol and of final growth of the preovulatory follicle did not differ between control and norgestomet-treated cows. However, there was a transient rise in FSH in association with weaning in control cows that did not occur in norgestomet-treated cows. In addition, mean LH increased more rapidly and mean oestradiol was higher during the 3 days immediately before the preovulatory surge of LH in the norgestomet-treated cows (P < 0.01).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Follicular development through preantral stages: signalling via growth factors.

Although gonadotrophins, particularly FSH, are the primary pituitary regulators of ovarian folliculogenesis, the involvement of ovarian-derived growth factors in follicular growth and maturation has become increasingly apparent over the past decade. Regulators of ovarian somatic cells can be broadly divided into (1) mitogenic factors, and (2) differentiation-induction factors, based on two primary cellular requirements, that is, proliferation and differentiation that are fundamental to folliculogenesis and ovarian functions. In this article, direct as well as indirect evidence is presented that highlights the roles of epidermal growth factor--a mitogen, and transforming growth factor beta--a differentiation-induction factor, in modulating gonadotrophin action in the ovary, particularly in the preantral follicles. An exquisitely timed and regionalized expression of these two types of peptide factor, along with their membrane receptors, may determine the differential success of follicle development, hence, allowing selection of the best oocytes for fertilization and subsequent development.

Animals↗

Effects of treatment with LH and FSH between 8 and 12 weeks of age on ovarian follicular development and puberty in heifers.

The transient increase in gonadotrophin secretion, seen in heifer calves between 6 and 20 wk of age, may be critical for early ovarian follicular growth and for initiation of sexual maturation. We treated heifers with either 3 mg of bLH (n = 5; sc) or 4 mg sc of bFSH (n = 5; sc), every other day, between 8 and 12 wk of age. During the first 17 d of treatment, bovine gonadotrophins caused a reduction in maximum antral follicle size and in numbers of large antral follicles, compared to control heifers (P < 0.05). In 4 of 5 bLH treated heifers and 3 of 5 bFSH treated heifers, the emergence of a large dominant antral follicle was delayed (P < 0.05). At 34 wk of age, follicular dynamics did not differ among groups. Serum concentrations of estradiol were decreased in bFSH and bLH treated heifers at 35 wk of age and in bFSH treated heifers at 25 wk of age compared with that of the controls. Time of first ovulation was delayed in bFSH treated heifers compared with control heifers (P < 0.05;bFSH 59.0 +/- 1.2; control 51.4 +/- 1.8; bLH 56.2 +/- 2.5 wk of age). In summary, treatment of 8-wk old heifers with gonadotrophins every other day disrupted folliculogenesis over a 17-d period, delayed first ovulation (bFSH), and decreased ovarian estradiol production at 25 (bFSH) and 35 (bLH, bFSH) weeks of age.

Journal Article↗

Follicular development in the sheep after priming with PMSG.

The administration of PMSG to sheep early in the oestrous cycle (Days 2 or 3) results in the formation of follicular cysts of varied morphology by Days 8 or 9. These may persist for up to 14 days after injection. If PMSG is given on Day 5 or later approximately 50% of such cysts ovulate. However, when PMSG is given at the beginning of the cycle (Day 2 or 3), the membrana granulosa is lost from the majority of the cysts and the theca interna luteinizes. The major hormone secreted by such luteinized follicles is progesterone. The structure of the steroid-secreting cells of the follicles is similar to that of large luteal cells of granulosa cell origin in cyclic corpora lutea. It is suggested that under suitable luteinizing conditions thecal cells may acquire many of the characteristics of granulosa cells.

Animals↗

The fine structure of the cumulus oophorus during follicular development in sheep.

The cumulus and membrana granulosa of non-atretic ovarian follicles from primordial up to a stage shortly before ovulation were studied by electron microscopy. The follicular cells of primordial follicles were undifferentiated and rested on a thick basal lamina. In secondary follicles the endoplasmic reticulum had proliferated forming an anastomosing network. In early antral and antral follicles (0.5--2.0 mm dia.) the ER was composed of short cisternae, the mitochondria had elongated and gap junctions were first observed. In late antral follicles (3.0--5.9 mm dia.) gap junctions were frequent. In the cumulus the glycogen was associated with electron lucent areas whereas in the granulosa it was invariably associated with membranes. In large antral follicles large membrane bound bodies were present in the basal cells of the cumulus. At early oestrus a distinctive mitochondrial morphology was noted in the granulosa but not elsewhere in the follicles. At mid oestrus numerous annular nexuses were present in the granulosa but not in the cumulus. At late oestrus numerous lipid droplets were formed in both cumulus and granulosa, the boundary with theca interna became indistinct and the basal lamina became incomplete.

Animals↗

Effect of GnRH antagonists treatment on gonadotrophin secretion, follicular development and inhibin A secretion in goats.

The aim of this study was to determine, for goats, the effects of daily doses of GnRH antagonist on ovarian endocrine and follicular function. Ten does were given 45 mg FGA intravaginal sponges and then five were treated with daily injections of 0.5mg of the GnRH antagonist Teverelix for 11 days from 2 days after the day of sponge insertion, while five does acted as controls. Pituitary activity was monitored by measuring plasma FSH and LH daily from 2 days before the first GnRH injection to Day 12. Follicular activity was determined by ultrasonographic monitoring and by assessing plasma inhibin A levels during the same period. In treated does, the FSH levels decreased linearly (0.8 +/- 0.1 ng/ml to 0.5 +/- 0.1 ng/ml, P < 0.01) and remained lower than the mean concentration in control goats (0.8 +/- 0.1 ng/ml, P < 0.005). LH levels were also lower during the period of antagonist treatment (0.6 +/- 0.2 ng/ml versus 0.4 +/- 0.1 ng/ml, P < 0.0005). During GnRH antagonist treatment, there was a significant decrease in the number of large follicles (> or = 6 mm) from Day 3 of treatment (1.2 +/- 0.6, P < 0.0001), with no large follicles from Day 9. The number of medium follicles (4-5 mm in size) also decrease during the period of treatment (4.2 +/- 0.7 to 1.0 +/- 0.6, P < 0.0001), leading to a significant decrease in inhibin A levels when compared to the control (143.7 +/- 31.3 pg/ml versus 65.2 +/- 19.1 pg/ml, P < 0.00005). In contrast, the number of small follicles (2-3 mm) increased in treated goats from Day 4 of treatment (9.6 +/- 2.9 to 20.2 +/- 6.3, P < 0.005). Such data indicate that GnRH antagonist reduced plasma levels of FSH and LH with suppression of the growth of large dominant ovarian follicles and a two-fold increase in number of smaller follicles. The results confirm that GnRH antagonist treatment can be used in goats to control gonadotrophin secretion and ovarian follicle growth in superovulatory regimes.

Administration, Intravaginal↗

Hormonal regulation of natriuretic peptide system during induced ovarian follicular development in the rat.

All components of the natriuretic peptide (NP) system have been found in the ovary. The purpose of this study was to determine the hormonal regulation of the NP system during follicular growth and ovulation induced by gonadotropins eCG and hCG. Ovarian membrane binding, before and after treatment, revealed the presence of guanylyl cyclase-type receptors exclusively. Equine CG treatment increased Bmax from 225 +/- 50 fmol/mg protein in control animals to 354 +/- 51 fmol/mg protein, and additional hCG treatment increased it further to 492 +/- 130 fmol/mg protein (p < 0.05), without changing receptor affinity. The increased binding was consistent with increased ability of atrial natriuretic peptide (ANP) to activate guanylyl cyclase in the ovarian cells obtained from hormone-treated animals. In confirmation, autoradiography of 125I-tyroCNP and 125I-ANP binding to the rat ovary showed that both guanylyl cyclase GC-A and GC-B receptor subtypes are localized to the granulosa cells of antral follicles. Quantitative analysis of GC-A and GC-B receptors by reverse transcription-polymerase chain reaction showed that the expression level of both receptors started to increase at 2 h and reached maximal levels at 6 h following eCG treatment. Increased levels of GC-B mRNA were also observed 12 h after eCG injection. At 24 and 48 h the receptor levels were below basal. Stimulation of NP receptors by eCG was paralleled by activation of both ovarian ANP and C-type natriuretic peptide (CNP) gene expression. ANP mRNA increased as early as 1 h after eCG injection and remained elevated up to 6 h. CNP mRNA increased at 2 h after eCG injection, peaked (5-fold) at 6 h, and remained elevated 48 h later, a stage at which follicular maturation continues. Incubation of ovaries with ANP significantly decreased eCG-induced estradiol level, indicating the functionality of the ovarian NP system. These results implicate the NP system in the induction and maintenance of fluid balance in the rapidly developing ovarian follicle.

Animals↗

The impact of previous danazol treatment on circulating hormone levels, follicular development and oocyte maturity in minimal-mild endometriosis.

The purpose of this study was to assess the impact of previous danazol treatment on peripheral endocrinology, folliculogenesis, oocyte maturity and follicular fluid endocrinology in women with minimal-mild endometriosis in a spontaneous menstrual cycle. A group of 10 women previously treated with danazol (Group A) and another group of 10 women whose endometriosis was left untreated (Group B) were studied. A group of 10 women with tubal infertility acted as a Control. Circulating hormone levels and characteristics of an endogenous luteinizing hormone (LH) surge were studied. A diagnostic laparoscopy was performed 32 h after the onset of an endogenous LH surge in all women to undertake follicular aspiration. There were no significant differences in oocyte maturity, fertilisation and cleavage rate among women studied in three study groups. The presence of peritoneal endometriosis did not affect the steroidogenic potential of the granulosa cells, and the endocrine milieu of the pre-ovulatory oocyte was comparable in all study groups.

Adult↗

Effect of long-term and short-term progestagen treatment on follicular development and pregnancy rate in cyclic ewes.

The aim of this study was to evaluate the effect of the length of a progestagen treatment (12 d vs. 6 d) on follicular dynamics, estrus synchronization and pregnancy rate using medroxyprogesterone acetate (MAP) with or without an eCG dose at the end of MAP treatment. One hundred sixty Polwarth ewes were divided into four equal groups: long-term treated (LT, n=40); short-term treated (ST, n=40); long-term treated plus eCG (LTeCG, n=40); and short-term treated plus eCG (STeCG, n=40). Five ewes of each group were separated to undergo daily transrectal ultrasonography, and blood samples were taken for hormone determination. Until 96 h after sponge withdrawal the number of ewes in estrus was higher in both long-term-treated groups than in both short-term-treated groups but at the end of the observational period (144 h) no significant differences were found among groups. The pregnancy rate was higher in the ST group (87%) than in the other groups (LT, 63%; LTeCG, 67%; and STeCG, 58%; P< or =0.03). The ovulatory follicle emerged before sponge withdrawal in long-term-treated ewes (-3.8+/-0.4 d and -2.2+/-0.8 d for LT and LTeCG, respectively), whereas in short-term-treated ewes it emerges around sponge removal (0.4+/-1.1 d and 0.5+/-0.5 d for ST and STeCG, respectively; P< or =0.01). The ovulatory follicle in the LT group had a longer lifespan and attained a larger (P< or =0.05) maximum diameter than in the ST group. We conclude: a) that the lower pregnancy rate observed after long-term progestagen treatment was related to a slower follicular turnover that promoted the ovulation of persistent dominant follicles; (b) that short-term treatment resulted in a higher pregnancy rate probably due to the ovulation of newly recruited growing follicles; and (c) treatment with eCG had no advantage in association with long-term treatment and had a deleterious effect in combination with short-term treatment with MAP.

Administration, Intravaginal↗

Effect of anaesthesia on ovarian follicular development and ovulation in the sheep subsequent to prostaglandin-induced luteolysis.

Prostaglandin analogues were used to induce luteal regression simultaneously in a number of ewes, thereby synchronizing the final stages of follicular maturation in these animals. Some of the ewes were anaesthetized for 24 h immediately after the injection of prostaglandin (experiment 1), and others for 15 h, starting 24 h after the injection of prostaglandin (experiment 2). In both experiments administration of anaesthetic significantly delayed the onset of oestrus and the time of ovulation relative to prostaglandin-treated control animals. The results from assays of blood samples collected at regular intervals in experiment 1 indicated that the preovulatory peak in the concentration of LH and the periovulatory changes in the concentration of FSH were similarly delayed and that during anaesthesia the level of LH was significantly reduced. It is suggested that the reduced level of LH, which probably resulted from a reduction in the secretion of releasing factor due to anaesthesia, failed to support oestrogen production by the Graafian follicle(s), thereby delaying the occurrence of oestrus and ovulation.

Anesthesia↗